AB0104 CHANGES IIN SECRETION OF Th17 EFFECTOR CYTOKINES IN SYNOVIAL FLUID CELL CULTURES BY THE ATOPIC DERMATITIS BIOLOGIC DUPILUMAB (ANTI-IL-4 RECEPTOR ALPHA)
Authors: M. P. Hundahl, S. Lomholt, T. W. Kragstrup
Journal: Annals of the Rheumatic Diseases (2022)

Background Inflammatory arthritis and enthesitis has been documented as adverse events in patients with atopic dermatitis treated with dupilumab targeting the interleukin 4 (IL-4) receptor alpha subunit (1,2). The disease mechanisms underlying atopic dermatitis are primarily driven by a substantiated Th2 response. Therefore, blockade of IL-4 and IL-13 signaling rationally inhibits Th2-mediated atopy. However, IL-4 and IL-13 are also well-known suppressors Th17 cells. Therefore, we hypothesize that dupilumab-induced arthritis could be caused by increased production of Th17 effector cytokines. Objectives Here, we tested the effect of dupilumab on in vivo activated peripheral blood and synovial fluid mononuclear cells from patients with active inflammatory arthritis. Methods We used peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) from patients with immune mediated inflammatory arthritis (juvenile idiopathic arthritis, spondylarthritis, polyarthritis, psoriatic arthritis, and arthritis with intestinal bowel disease (IBD), n=8). PBMCs and SFMCs were either unstimulated or stimulated with CD3/Cd28 beads, and then cultured with three different concentrations of dupilumab (0.2 μg/mL, 1.0 μg/mL or 5.0 μg/mL). Supernatants were analyzed by the two MSD Multi-Spot Assay System in the Proinflammatory Panel 1 (INF-γ, IL-1β, IL-2, Il-4, IL-6, IL-8, IL-10, IL-12p70, IL-13 and TNF-α) and the Th17 Panel 1 (IL-17A, IL-21, IL-22, IL-23, IL-27, IL-31 and CCL20). All data were transformed to ratios by dividing the value of the samples treated with dupilumab with the value of untreated samples. Data were then analyzed with repeated measures one-way ANOVA or the paired t-test depending on the number of groups. P-values &lt;0.05 were considered statistically significant. Results In SFMCs not cultured with CD3/CD28 beads, dupilumab decreased the production of IL-17A, IL-22, CCL20 and IL-1β (P-values: 0.0007, 0.0002, 0. 0025 and 0.0432 respectively). IL-6 was decreased by dupilumab in both PBMCs and SFMCs not cultured with CD3/CD28 beads (P-value = 0.013 and P-value = 0.001). The concentration of cytokines measured in supernatants from the PBMCs cultured without CD3/CD28 were low in both the Proinflammatory Panel and the Th17 panel. Interestingly, the response to increasing concentrations of dupilumab in PBMCs and SFMCs pre-conditioned with CD3/CD28 showed a biphasic curve. Thus, the low concentration of dupilumab (0.2 μg/mL) resulted in increased concentration while the higher concentrations of dupilumab (5.0 μg/mL) decreased the concentration. This was particularly seen in CD3/CD28-stimulated SFMC cultures for IL-17A, IL-22, IL-4, IL-13, IL-31, IL-10 and IL-1β (P-value for UT vs. 0.2 µg/mL: 0.0117, 0.0379, 0.0052, 0.0002, 0.0456, 0.0003 and 0.0484) In CD3/CD28-stimulated PBMC cultures, the biphasic response curve was seen for TNF-α, IFN-γ, IL-2 and IL-10 (P-value for UT vs. 1.0 µg/mL: 0.4978, 0.3922, 0.0373 and 0.0008). Conclusion This data contradicts the hypothesis and indicates that dupilumab does not increase the production of Th17 related cytokines. However, the increase in proinflammatory cytokines seen with the low concentrations of dupilumab suggest that partial blockade of the IL-4 receptor alpha under some conditions could lead to augmented inflammation. References [1]Willsmore, Z.N. , et al. Development of inflammatory arthritis and enthesitis in patients on dupilumab: a case series. Br J Dermatol 181 , 1068-1070 (2019). [2]Ishibashi, M., Honda, T., Tabuchi, Y. &amp; Kabashima, K. Polyenthesitis during treatment with dupilumab for atopic dermatitis. J Eur Acad Dermatol Venereol 34 , e319-e321 (2020). Figure 1. The biphasic response to dupilumab treatment shown for the analytes IL-17A and IL-22. An increase is seen in UT vs. 0.2 μg/mL with a P-value of 0.0117 and 0.0379. Disclosure of Interests Malthe Pallesgaard Hundahl: None declared, Søren Lomholt: None declared, Tue Wenzel Kragstrup Shareholder of: Co-founder and clinical developer in iBio tech ApS, Speakers bureau: Speaking fees from Pfizer, Bristol-Myers Squibb, Eli Lilly, Novartis, UCB, and Abbvie, Consultant of: Consultancy fees from Bristol-Myers Squibb and Gilead, Grant/research support from: Research grants from Gilead